Identification of a Novel Linear B-Cell Epitope in the M Protein of Avian Infectious Bronchitis Coronaviruses

Junji Xing,Shengwang Liu,Zongxi Han,Yuhao Shao,Huixin Li,Xiangang Kong 한국미생물학회 2009 The journal of microbiology Vol.47 No.5

This report describes the identification of a novel linear B-cell epitope at the C-terminus of the membrane (M) protein of avian infectious bronchitis virus (IBV). A monoclonal antibody (MAb) (designated as 15E2) against the IBV M protein was prepared and a series of 14 partially-overlapping fragments of the IBV M gene were expressed with a GST tag. These peptides were subjected to enzyme-linked immunosorbent assay (ELISA) and western blotting analysis using MAb 15E2 to identify the epitope. A linear motif, 199FATFVYAK206, which was located at the C-terminus of the M protein, was identified by MAb 15E2. ELISA and western blotting also showed that this epitope could be recognized by IBV-positive serum from chicken. Given that 15E2 showed reactivity with the 199FATFVYAK206 motif, expressed as a GST fusion protein, in both western blotting and in an ELISA, we proposed that this motif represented a linear B-cell epitope of the M protein. The 199FATFVYAK206 motif was the minimal requirement for reactivity as demonstrated by analysis of the reactivity of 15E2 with several truncated peptides that were derived from the motif. Alignment and comparison of the 15E2-defined epitope sequence with the sequences of other coronaviruses indicated that the epitope is well conserved among chicken and turkey coronaviruses. The identified epitope should be useful in clinical applications and as a tool for the further study of the structure and function of the M protein of IBV.

Identification and Expression Patterns of fvexpl1, an Expansin-Like Protein-Encoding Gene, Suggest an Auxiliary Role in the Stipe Morphogenesis of Flammulina velutipes

( Qianhui Huang ),( Xing Han ),( Irum Mukhtar ),( Lingling Gao ),( Rongmei Huang ),( Liping Fu ),( Junjie Yan ),( Yongxin Tao ),( Bingzhi Chen ),( Baogui Xie ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.4

Expansins are cell wall proteins that mediate cell wall loosening and promote specific tissue and organ morphogenesis in plants and in some microorganisms. Unlike plant expansins, the biological functions of fungal expansin-like proteins have rarely been discussed. In the present study, an expansin-like protein-encoding fvexpl1 gene, was identified from Flammulina velutipes by using local BLAST. It consisted of five exons with a total length of 822 bp. The deduced protein FVEXPL1 contained 274 amino acids with a predicted molecular mass and isoelectric point of 28,589 Da and pH 4.93, respectively. The first 19 amino acids from the N terminal are the signal peptide. Phylogenetic analysis and multiple protein alignment indicated FVEXPL1 was an expansin-like protein. The expression level of fvexpl1 gene in the stipe was significantly higher than that in the mycelia, primordia, and cap. However, the expression level of fvexpl1 gene was significantly higher in the fast elongation region of the stipe as compared with the slow elongation region. Expression analysis indicated that fvexpl1 gene might have an auxiliary role in the stipe morphogenesis of F. velutipes.

LDMOS with Variable-k Dielectric for Improved Breakdown Voltage and Specific On-resistance

Songnan Guo,Junji Cheng,Xing Bi Chen 대한전자공학회 2019 Journal of semiconductor technology and science Vol.19 No.5

A silicon-on-insulator (SOI) lateral double-diffused MOSFET (LDMOS) with variable-k dielectric is proposed, which features three-section high-k dielectrics placed over the drift region. Beneficial from the modulation, by high-k dielectrics, on electric fluxes in the drift region, the electric field strength is locally enhanced, and has a more uniform distribution along the surface of the drift region, which an increment of the breakdown voltage (BV) ascribes to. Moreover, due to the electric fluxes prefer to flow towards and through a high-k dielectric, an increase in the dose of the drift region is thus possible to reduce the specific on-resistance (Ron,sp), since the extra electric fluxes caused by those increased impurities would flow, upwards, into the high-k dielectric, and barely make contributions to the x-component of the surface electric field, which indicates that the BV would not be affected. Based on a comparison with the conventional LDMOS, the BV of the proposed LDMOS increased by 31% with a decrease of 13% in the Ron,sp. The figure of merit (FOM) increased approximately 2 times.

Molecular Cloning and Expression of Grass Carp MyoD in Yeast Pichia pastoris

Wang, Lixin,Bai, Junjie,Luo, Jianren,Chen, Hong,Ye, Xing,Jian, Qing,Lao, Haihua Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.1

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

Molecular mapping of four blast resistance genes using recombinant inbred lines of 93-11 and nipponbare

Hongmei Yang,Melissa H Jia,Yulin Jia,Junjie Xing,Venu, R-C,Maria Bellizzi,Longping Yuan,Zhilong Wang,Chuanqing Sun,Guo-Liang Wang 한국식물학회 2013 Journal of Plant Biology Vol.56 No.2

Molecular mapping of new blast resistance genes is important for developing resistant rice cultivars using marker-assisted selection. In this study, 259 recombinant inbred lines (RILs) were developed from a cross between Nipponbare and 93-11, and were used to construct a 1165.8-cM linkage map with 131 polymorphic simple sequence repeat (SSR) markers. Four major quantitative trait loci (QTLs) for resistance to six isolates of Magnaporthe oryzae were identified: qPi93-1, qPi93-2, qPi93-3, and qPiN-1. For the three genes identified in 93-11, qPi93-1 is linked with SSR marker RM116 on the short arm of chromosome 11 and explains 33% of the phenotypic variation in resistance to isolate CHE86. qPi93-2 is linked with SSR marker RM224 on the long arm of chromosome 11 and accounts for 31% and 25% of the phenotypic variation in resistance to isolates 162-8B and ARB50, respectively. qPi93-3 is linked with SSR marker RM7102 on chromosome 12 and explains 16%, 53%, and 28% of the phenotypic variation in resistance to isolates CHE86, ARB52, and ARB94, respectively. QTL qPiN-1 from Nipponbare is associated with SSR marker RM302 on chromosome 1 and accounts for 34% of the phenotypic variation in resistance to isolate PO6-6. These new genes can be used to develop new varieties with blast resistance via marker-aided selection and to explore the molecular mechanism of rice blast resistance.

Identification and Characterization of Two New S-Adenosylmethionine- Dependent Methyltransferase Encoding Genes Suggested Their Involvement in Stipe Elongation of Flammulina velutipes

( Qianhui Huang ),( Irum Mukhtar ),( Yelin Zhang ),( Zhongyang Wei ),( Xing Han ),( Rongmei Huang ),( Junjie Yan ),( Baogui Xie ) 한국균학회 2019 Mycobiology Vol.47 No.4

Two new SAM-dependent methyltransferase encoding genes (fvsmt1 and fvsmt2) were identified from the genome of Flammulina velutipes. In order to make a comprehensive characterization of both genes, we performed in silico analysis of both genes and used qRT-PCR to reveal their expression patterns during the development of F. velutipes. There are 4 and 6 exons with total length of 693 and 978 bp in fvsmt2 and fvsmt1, respectively. The deduced proteins, i.e., FVSMT1 and FVSMT2 contained 325 and 230 amino acids with molecular weight 36297 and 24894 Da, respectively. Both proteins contained a SAM-dependent catalytic domain with signature motifs (I, p-I, II, and III) defining the SAM fold. SAM-dependent catalytic domain is located either in the middle or at the N-terminal of FVSMT2 and FVSMT1, respectively. Alignment and phylogenic analysis showed that FVSMT1 is a homolog to a protein- arginine omega-N-methyltransferase, while FVSMT2 is of cinnamoyl CoA O-methyltransferase type and predicted subcellular locations of these proteins are mitochondria and cytoplasm, respectively. qRT-PCR showed that fvsmt1 and fvsmt2 expression was regulated in different developmental stages. The maximum expression levels of fvsmt1 and fvsmt2 were observed in stipe elongation, while no difference was found in mycelium and pileus. These results positively demonstrate that both the methyltransferase encoding genes are involved in the stipe elongation of F. velutipes.

Optimal Two-Section Layouts for the Two-Dimensional Cutting Problem

( Jun Ji ),( Dun-hua Huang ),( Fei-fei Xing ),( Yao-dong Cui ) 한국정보처리학회 2021 Journal of information processing systems Vol.17 No.2

When generating layout schemes, both the material usage and practicality of the cutting process should be considered. This paper presents a two-section algorithm for generating guillotine-cutting schemes of rectangular blanks. It simplifies the cutting process by allowing only one size of blanks to appear in any rectangular block. The algorithm uses an implicit enumeration and a linear programming optimal cutting scheme to maximize the material usage. The algorithm was tested on some benchmark problems in the literature, and compared with the three types of layout scheme algorithm. The experimental results show that the algorithm is effective both in computation time and in material usage.